Lipids

, Volume 47, Issue 12, pp 1221–1229

An Improved Method for Analysis of Glucosylceramide Species Having cis-8 and trans-8 Isomers of Sphingoid Bases by LC–MS/MS

  • Hiroyuki Imai
  • Hideyasu Hattori
  • Masayuki Watanabe
Methods
  • 657 Downloads

Abstract

High-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC–ESI–MS/MS) approaches have enabled high selectivity and sensitivity for the identification and quantification of glucosylceramide molecular species. Here we demonstrate that HPLC–ESI–MS/MS is an efficient method for characterizing plant glucosylceramide species having the cis-8 and trans-8 isomers of sphingoid bases. Complete baseline separation was achieved using a high-carbon-content octadecylsilyl column and a simple binary gradient comprising methanol and water. The result of 2-hydroxy fatty acid composition achieved by HPLC–ESI–MS/MS was compared with that achieved by gas chromatography with flame ionization detection (GC–FID), indicating that the two methods yield similar molar compositions. The current method should be applicable to seeking the active components of glucosylceramide species from plant materials in response to biological challenges.

Keywords

Glucosylceramides Glycosphingolipids Liquid chromatography–electrospray ionization tandem mass spectrometry Plant lipids Sphingoid bases 

Abbreviations

d18:2(t4,c8)

trans-4,cis-8-Sphingadienine

d18:2(t4,t8)

trans-4,trans-8-Sphingadienine

ESI

Electrospray ionization

FID

Flame ionization detection

GC

Gas chromatography

Glc

Glucose

HPLC

High-performance liquid chromatography

LC

Liquid chromatography

MS/MS

Tandem mass spectrometry

ODS

Octadecylsilyl

t18:1(c8)

4-Hydroxy-cis-8-sphingenine

t18:1(t8)

4-Hydroxy-trans-8-sphingenine

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Copyright information

© AOCS 2012

Authors and Affiliations

  • Hiroyuki Imai
    • 1
  • Hideyasu Hattori
    • 2
  • Masayuki Watanabe
    • 1
  1. 1.Department of Biology, Graduate School of Natural ScienceKonan UniversityKobeJapan
  2. 2.Development and Marketing Group, Frontier Science Business DivisionShiseido Co., Ltd.KyotoJapan

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